DE102008017103A1 - Detergents and cleaning agents containing proteases from Xanthomonas - Google Patents

Abstract

The invention relates to detergents and cleaners containing a protease from a bacterium of the genus Xanthomonas and these proteases themselves. Furthermore, the invention relates to purification processes in which these agents are used as well as uses of these agents. Furthermore, the invention relates to the preparation and use of these proteases themselves.

Description

The The invention relates to detergents and cleaners containing a protease from a bacterium of the genus Xanthomonas included as well as these Proteases themselves. Furthermore, the invention relates to purification processes, in which these funds are used and uses of these Medium. Furthermore, the invention relates to the preparation and use of these proteases themselves.

For Detergents and cleaners are so far preferred proteases from Used subtilisin type. The in the from the state of the art originate from known detergents or cleaning agents used proteases either originally from microorganisms, for example genera Bacillus, Streptomyces, Humicola, or Pseudomonas, and / or are used according to known biotechnological processes produced by suitable microorganisms, for example by transgenic expression hosts of the genera Bacillus or by filamentous Mushrooms.

Examples thereof are the subtilisins BPN 'and Carlsberg, the protease PB92, the subtilisins 147 and 309, the alkaline protease from Bacillus lentus, subtilisin DY and the enzymes thermitase, proteinase K and the subtilases, but not the subtilisins in the narrower sense Proteases TW3 and TW7. Subtilisin Carlsberg in a developed form under the trade names Alcalase ^® from Novozymes A / S, Bagsvaerd, Denmark. The subtilisins 147 and 309 are sold under the trade names Esperase ^®, or Savinase ^® from Novozymes. From the protease from Bacillus lentus DSM 5483 ( WO 91/02792 A1 ) derive from the name under the name BLAP ^® variants, which in particular in WO 92/21760 A1 . WO 95/23221 A1 . WO 02/088340 A2 and WO 03/038082 A2 to be discribed. Other useful proteases from various Bacillus species are found in the patent applications WO 03/054185 A1 . WO 03/056017 A2 . WO 03/055974 A2 and WO 03/054184 A1 out.

Natural proteases formed by microorganisms, which are isolatable from natural habitats and can be used in detergents and cleaners, for example, go from the applications WO 03/054185 A1 (from Bacillus gibsonii (DSM 14391)), WO 03/056017 A2 (from Bacillus sp. (DSM 14390)), WO 03/055974 A2 (from Bacillus sp. (DSM 14392)) and WO 03/054184 A1 (from Bacillus gibsonii (DSM 14393)). All of these applications also disclose corresponding detergents and cleaners containing these proteases.

Out the prior art is also known that bacteria of the genus Xanthomonas produce enzymes that have a proteolytic activity have. These enzymes are often associated with diseases of plants associated bacteria of the genus Xanthomonas as Cause plant pathogens.

Thus, Dow et al. in two publications ( Applied and Environmental Microbiology 1990, 2994-2998 such as Applied and Environmental Microbiology 1993, 3996-4003 ) the proteases PRT1 and PRT2 from Xanthomonas campestris against the background of plant pathogenesis.

In Chang-Ho Lee et al. (Jour, Microbiol., 1995, 115-119) an alkaline protease disclosed by Xanthomonas sp. YL-37 is formed. In addition to the general statement that proteolytic enzymes are used per se in laundry products, no detergents or cleaners containing a protease from Xanthomonas are disclosed.

Out The prior art are thus no detergents and cleaners known in which a protease from a bacterium of the genus Xanthomonas is used.

One Disadvantage of the hitherto used in detergents and cleaners Proteases of the prior art is that they are often not have satisfactory proteolytic activity and the Mitttel containing them therefore based on proteinaceous Soiling does not have a satisfactory cleaning performance. In particular, this is the case at low temperatures, for example between 20 ° C and 60 ° C, so that they contain Detergents or cleaning agents in this temperature range no optimal Show cleaning performance. Thus, there is a need to have new proteases, especially new microbial proteases, for use to be found in detergents and cleaners and accordingly new ones To provide detergents and cleaners containing such proteases contain.

It is therefore an object of the present invention to provide detergents which exhibit improved removal of proteinaceous residues with respect to at least one soiling, preferably with respect to multiple soils. In particular, should the washing or Detergents show improved removal of proteinaceous residues with respect to at least one soiling, preferably with respect to multiple soils, at low temperatures, especially in a temperature range between 20 and 60 ° C.

A Another object of the present invention is precisely such protease enzymes to be provided for use in accordance with the invention Detergents and cleaning agents are suitable and are characterized by a good, preferred by an improved proteolytic activity with respect to at least one soiling, preferably with respect to on several stains, when they wash or in a wash Detergents are used, in particular in the above mentioned temperature range.

These The object is achieved according to the invention The teaching of claim 1 by the provision of washing or cleaning agents, the one protease that occurs naturally in a bacterium of the genus Xanthomonas is present and at least one other detergent ingredient include. Surprisingly, it was found that Proteases derived from a bacterium of the genus Xanthomonas, advantageous in washing or cleaning agents according to the invention can be used and in washing according to the invention or cleaning agents, especially at low temperatures, especially in a temperature range between 20 and 60 ° C, an improved removal of protease-sensitive residues, for example Soiling on textiles or dishes.

a The invention thus provides washing or cleaning agents, the one protease that is in a bacterium of the genus Xanthomonas is naturally present and at least one other detergent ingredient include. In a preferred embodiment is a Such detergent or cleaning agent, characterized in that the bacterium is selected from the group consisting of Xanthomonas albilineans, Xanthomonas alfalfae, Xanthomonas alfalfae subsp. alfalfae, Xanthomonas alfalfae subsp. citrumelonis, Xanthomonas ampelina (Xylophilus ampelinus), Xanthomonas arboricola, Xanthomonas axonopodis, Xanthomonas bromi, Xanthomonas campestris, Xanthomonas cassavae, Xanthomonas citri, Xanthomonas citri subsp. citri, Xanthomonas citri subsp. malvacearum, Xanthomonas codiaei, Xanthomonas cucurbitae, Xanthomonas cynarae, Xanthomonas euvesicatoria, Xanthomonas fragariae, Xanthomonas fuscans, Xanthomonas fuscans subsp. aurantifolii, Xanthomonas fuscans subsp. fuscans, Xanthomonas gardneri, Xanthomonas hortorum, Xanthomonas hyacinthi, Xanthomonas melonis, Xanthomonas oryzae, Xanthomonas perforans, Xanthomonas phaseoli, Xanthomonas pisi, Xanthomonas populi, Xanthomonas sacchari, Xanthomonas theicola, Xanthomonas translucens, Xanthomonas vasicola, Xanthomonas vesicatoria. It has It has been found that especially these Xanthomonas species proteases expressing those in washing or Detergents show corresponding advantages.

naturally present in this context means that the protease has a bacterial protease that is isolated from the bacterium can. Therefore, in particular, such proteases are not included, with the help of genetic engineering methods in an inventive Bacterial strain were introduced and expressed by this recombinant become. Proteases known from the prior art which are prepared with the aid of produces a bacterial strain according to the invention be - d. H. for the bacteria of the genus Xanthomonas the production organism on the basis of the introduction of the the protease-encoding gene in this organism by means of genetic engineering Method is - are therefore not the subject of the invention.

The The classical procedure for obtaining the enzymes is obvious to the person skilled in the art known in the field of enzyme technology and consists of the microorganism-containing Samples from, for example, natural habitats and under conditions deemed suitable - for Example in alkaline medium - to cultivate. To this One obtains enrichment cultures of microorganisms, the desired enzymes, in the present case the Protease enzymes, which are present under the relevant conditions are active. This will then, for example, over Ausplattieren on protein-containing agar plates and measuring the lysis court formed the microorganisms selected with the most powerful enzymes and / or the genes encoding them identified and cloned.

Such an approach is described, for example, in the textbook "Alkalophilic Microorganisms. A new microbial world "by K. Horikoshi and T. Akiba (1982), Japan Scientific Societies Press, Springer-Verlag, New York, Heidelberg, Berlin, ISBN 0-387-10924-2, Chapter 2, pp. 9-26 described. For example, too WO 00/24882 A1 discloses a method for producing a gene bank obtained by culturing and thus enriching microorganism-containing samples from any habitat, for example, the rumen, under the conditions of interest, and isolating and cloning nucleic acids of interest thereto.

however it is by no means such a protease that naturally in a microorganism, especially a bacterium is, is also advantageously used in a detergent or cleaning agent, d. H. in these agents a satisfactory proteolytic Activity, for example, when using a containing detergent or cleaning agent in a satisfactory Removal of proteinaceous soiling results. thats why it is all the more surprising that in bacteria of the genus Xanthomonas, in particular in the Xanthomonas species described above, Proteases are present that are generally for use in detergents and cleaners according to the invention and in particular when using a wash containing them. and detergent a satisfactory removal of proteinaceous Cause soiling. Bacteria of this bacterial genus - and in particular the aforementioned species - possess naturally, without genetic or molecular biology Modification, proteolytic enzymes that are among the most demanding Conditions of use of detergents and cleaners sufficient have high proteolytic activity to proteinaceous Soiling under the conditions of use of the washing or cleaning agent dismantle. Demanding conditions of use for detergents and cleaners arise in particular due to the presence of one or several other ingredients in such agents and in the they formed wash liquor during the washing process like Bleaching agents, bleach activators, surfactants, builders and / or builders due to the pH of such agents and those formed by them Washing liquor during the washing process and / or on reason the ionic strength and / or the temperature of the wash liquor during the washing process.

Within the genus Xanthomonas, some strains have been found to be particularly advantageous. These naturally contain proteases which are particularly suitable for use in detergents and cleaners according to the invention. In a preferred embodiment of the invention, the agent is characterized in that the bacterium is selected from the group consisting of Xanthomonas axonopodis DSM 3585, Xanthomonas axonopodis pvar. Begoniae DSM 50850, Xanthomonas axonopodis pvar. malvacearum DSM 1220, Xanthomonas campestris DSM 1050 (NCPPB 1929), Xanthomonas campestris DSM 19000, Xanthomonas campestris pvar. campestris DSM 3586T, Xanthomonas campestris pvar. pelargonii DSM 50857. These strains are deposited and publicly available at the German Collection of Microorganisms and Cell Cultures (DSMZ), Inhoffenstr. 7b, 38124 Braunschweig, Germany ( www.dsmz.de ).

It was found to be a detergent with a According to the invention preferred protease increased Has performance over a protease-free agent and a very good cleaning performance in terms of protease-sensitive Soiling reaches.

To This subject matter includes all conceivable washing or detergent types, both concentrates and undiluted means to be used, for commercial scale use, in the washing machine or in hand washing respectively -cleaning. These include, for example, detergents for Textiles, carpets or natural fibers for which the present invention, the term detergent is used. These include, for example, dishwashing detergents for dishwashers or manual dishwashing detergents or cleaner for hard surfaces like metal, Glass, porcelain, ceramics, tiles, stone, painted surfaces, Plastics, wood or leather; for such will be after the present invention uses the term cleaning agent. An inventive washing or cleaning agent can be both a means for bulk consumers or technical users as well as a product for the private consumer be. According to the invention, therefore, all types of appropriate detergents or cleaning agents in the protected area of the present invention, their applicability by adding an above-described inventive Protein is improved.

Prefers contains an agent according to the invention the protease in an amount of 2 μg to 20 mg, preferably from 5 μg to 17.5 mg, more preferably 20 μg to 15 mg and most preferably from 50 μg to 10 mg per g of the agent.

The washing or cleaning agents according to the invention, which can be in the form of homogeneous solutions or suspensions in particular as powdered solids, can in addition to the active ingredient used according to the invention - a protease from a bacterium of the genus Xanthomonas - in principle all known and customary in such agents Containing ingredients, wherein at least one further ingredient is present in the agent. The agents according to the invention may in particular be builders, surface-active surfactants, bleaches based on organic and / or inorganic peroxygen compounds, bleach activators, water-miscible organic solvents, enzymes, sequestering agents, electrolytes, pH regulators and other auxiliaries such as optical brighteners, graying inhibitors gates, foam regulators and dyes and fragrances and combinations thereof.

Especially a combination of a protease present in a bacterium of the genus Xanthomonas is naturally present with one or several other ingredient (s) of the agent proves to be advantageous because such a means improved cleaning performance resulting synergisms, in particular between the protease and the further ingredient. This means that the means an improved removal of stains, such as proteinaceous Stains, in comparison with a remedy, either contains only one of the two components, or even in comparison to the expected cleaning performance of an agent with both components due to the mere addition of the individual contributions of these two components for cleaning performance of the agent. In particular by the combination of a protease present in a bacterium of the genus Xanthomonas is naturally present with one of the surfactants and / or builders described below and / or Bleach will achieve such synergism.

The Agents according to the invention can be a surfactant or more surfactants, especially anionic surfactants, nonionic surfactants and their mixtures, but also cationic, zwitterionic and amphoteric surfactants come into question.

suitable Nonionic surfactants are, in particular, alkyl glycosides and ethoxylation and / or propoxylation products of alkyl glycosides or linear or branched alcohols each having 12 to 18 carbon atoms in the alkyl moiety and 3 to 20, preferably 4 to 10 alkyl ether groups. Farther are corresponding ethoxylation and / or propoxylation products of N-alkylamines, vicinal diols, fatty acid esters and fatty acid amides, with respect to the alkyl part said long-chain alcohol derivatives correspond, and of alkylphenols having 5 to 12 carbon atoms in the alkyl radical useful.

The nonionic surfactants used are preferably alkoxylated, advantageously ethoxylated, in particular primary, alcohols having preferably 8 to 18 carbon atoms and on average 1 to 12 moles of ethylene oxide (EO) per mole of alcohol, in which the alcohol radical can be linear or preferably methyl-branched in the 2-position or may contain linear and methyl-branched radicals in the mixture, as they are usually present in Oxoalkoholresten. In particular, however, alcohol ethoxylates with linear radicals of alcohols of native origin having 12 to 18 carbon atoms, for. From coconut, palm, tallow or oleyl alcohol, and on average from 2 to 8 EO per mole of alcohol. The preferred ethoxylated alcohols include, for example, C ₁₂ -C ₁₄ -alcohols with 3 EO or 4 EO, C ₉ -C ₁₁ -alcohols with 7 EO, C ₁₃ -C ₁₅ -alcohols with 3 EO, 5 EO, 7 EO or 8 EO, C ₁₂ -C ₁₈ -alcohols with 3 EO, 5 EO or 7 EO and mixtures of these, such as mixtures of C ₁₂ -C ₁₄ -alcohol with 3 EO and C ₁₂ -C ₁₈ -alcohol with 7 EO. The degrees of ethoxylation given represent statistical means which, for a particular product, may be an integer or a fractional number. Preferred alcohol ethoxylates have a narrow homolog distribution (narrow range ethoxylates, NRE). In addition to these nonionic surfactants, fatty alcohols with more than 12 EO can also be used. Examples include (tallow) fatty alcohols with 14 EO, 16 EO, 20 EO, 25 EO, 30 EO or 40 EO. In particular, agents for use in mechanical processes usually extremely low-foam compounds are used. These include preferably C ₁₂ -C ₁₈ -alkylpolyethylenglykol-polypropylene glycol ethers with in each case at to 8 mol ethylene oxide and propylene oxide units in the molecule. However, it is also possible to use other known low-foam nonionic surfactants, such as, for example, C ₁₂ -C ₁₈ -alkyl polyethylene glycol-polybutylene glycol ethers having in each case up to 8 mol of ethylene oxide and butylene oxide units in the molecule and end-capped alkylpolyalkylene glycol mixed ethers. Also particularly preferred are the hydroxyl-containing alkoxylated alcohols, as described in the European patent application EP 0 300 305 are described, so-called Hydroxymischether. The nonionic surfactants also include alkyl glycosides of the general formula RO (G) _x in which R is a primary straight-chain or methyl-branched, in particular 2-methyl-branched aliphatic radical having 8 to 22, preferably 12 to 18 carbon atoms and G represents a glycose unit having 5 or 6 C atoms, preferably glucose. The degree of oligomerization x, which indicates the distribution of monoglycosides and oligoglycosides, is an arbitrary number - which, as a variable to be determined analytically, may also assume fractional values - between 1 and 10; preferably x is 1.2 to 1.4. Also suitable are polyhydroxy fatty acid amides of the formula (III) in which R ¹ CO is an aliphatic acyl radical having 6 to 22 carbon atoms, R ^{2 is} hydrogen, an alkyl or hydroxyalkyl radical having 1 to 4 carbon atoms and [Z] is a linear or branched polyhydroxyalkyl radical having 3 to 10 carbon atoms and 3 to 10 hydroxyl groups:

in der R³ für einen linearen oder verzweigten Alkyl- oder Alkenylrest mit 7 bis 12 Kohlenstoffatomen, R⁴ für einen linearen, verzweigten oder cyclischen Alkylenrest oder einen Arylenrest mit 2 bis 8 Kohlenstoffatomen und R⁵ für einen linearen, verzweigten oder cyclischen Alkylrest oder einen Arylrest oder einen Oxy-Alkylrest mit 1 bis 8 Kohlenstoffatomen steht, wobei C₁-C₄-Alkyl- oder Phenylreste bevorzugt sind, und [Z] für einen linearen Polyhydroxyalkylrest, dessen Alkylkette mit mindestens zwei Hydroxylgruppen substituiert ist, oder alkoxylierte, vorzugsweise ethoxylierte oder propoxylierte Derivate dieses Restes steht. [Z] wird auch hier vorzugsweise durch reduktive Aminierung eines Zuckers wie Glucose, Fructose, Maltose, Lactose, Galactose, Mannose oder Xylose erhalten. Die N-Alkoxy- oder N-Aryloxy-substituierten Verbindungen können dann beispielsweise durch Umsetzung mit Fettsäuremethylestern in Gegenwart eines Alkoxids als Katalysator in die gewünschten Polyhydroxyfettsäureamide überführt werden. Eine weitere Klasse bevorzugt eingesetzter nichtionischer Tenside, die entweder als alleiniges nichtionisches Tensid oder in Kombination mit anderen nichtionischen Tensiden, insbesondere zusammen mit alkoxylierten Fettalkoholen und/oder Alkylglykosiden, eingesetzt werden, sind alkoxylierte, vorzugsweise ethoxylierte oder ethoxylierte und propoxylierte Fettsäurealkylester, vorzugsweise mit 1 bis 4 Kohlenstoffatomen in der Alkylkette, insbesondere Fettsäuremethylester. Auch nichtionische Tenside vom Typ der Aminoxide, beispielsweise N-Kokosalkyl-N,N-dimethylaminoxid und N-Talgalkyl-N,N-dihydroxyethylaminoxid, und der Fettsäurealkanolamide können geeignet sein. Die Menge dieser nichtionischen Tenside beträgt vorzugsweise nicht mehr als die der ethoxylierten Fettalkohole, insbesondere nicht mehr als die Hälfte davon. Als weitere Tenside kommen sogenannte Gemini-Tenside in Betracht. Hierunter werden im Allgemeinen solche Verbindungen verstanden, die zwei hydrophile Gruppen pro Molekül besitzen. Diese Gruppen sind in der Regel durch einen sogenannten ”Spacer” voneinander getrennt. Dieser Spacer ist in der Regel eine Kohlenstoffkette, die lang genug sein sollte, dass die hydrophilen Gruppen einen ausreichenden Abstand haben, damit sie unabhängig voneinander agieren können. Derartige Tenside zeichnen sich im Allgemeinen durch eine ungewöhnlich geringe kritische Micellkonzentration und die Fähigkeit, die Oberflächenspannung des Wassers stark zu reduzieren, aus. In Ausnahmefällen werden unter dem Ausdruck Gemini-Tenside nicht nur derartig ”dimere”, sondern auch entsprechend ”trimere” Tenside verstanden. Geeignete Gemini-Tenside sind beispielsweise sulfatierte Hydroxymischether oder Dimeralkohol-bis- und Trimeralkohol-tris-sulfate und -ethersulfate. Endgruppenverschlossene dimere und trimere Mischether zeichnen sich insbesondere durch ihre Bi- und Multifunktionalität aus. So besitzen die genannten endgruppenverschlossenen Tenside gute Netzeigenschaften und sind dabei schaumarm, so dass sie sich insbesondere für den Einsatz in maschinellen Wasch- oder Reinigungsverfahren eignen. Eingesetzt werden können aber auch Gemini-Polyhydroxyfettsäureamide oder Poly-Polyhydroxyfettsäureamide. Geeignet sind auch die Schwefelsäuremonoester der mit 1 bis 6 Mol Ethylenoxid ethoxylierten geradkettigen oder verzweigten C₇-C₂₁-Alkohole, wie 2-Methylverzweigte C₉-C₁₁-Alkohole mit im Durchschnitt 3,5 Mol Ethylenoxid (EO) oder C₁₂-C₁₈-Fettalkohole mit 1 bis 4 EO. Zu den bevorzugten Aniontensiden gehören auch die Salze der Alkylsulfobernsteinsäure, die auch als Sulfosuccinate oder als Sulfobernsteinsäureester bezeichnet werden, und die Monoester und/oder Diester der Sulfobernsteinsäure mit Alkoholen, vorzugsweise Fettalkoholen und insbesondere ethoxylierten Fettalkoholen darstellen. Bevorzugte Sulfosuccinate enthalten C₈- bis C₁₈-Fettalkoholreste oder Mischungen aus diesen. Insbesondere bevorzugte Sulfosuccinate enthalten einen Fettalkoholrest, der sich von ethoxylierten Fettalkoholen ableitet, die für sich betrachtet nichtionische Tenside darstellen. Dabei sind wiederum Sulfosuccinate, deren Fettalkohol-Reste sich von ethoxylierten Fettalkoholen mit eingeengter Homologenverteilung ableiten, besonders bevorzugt. Ebenso ist es auch möglich, Alk(en)ylbernsteinsäure mit vorzugsweise 8 bis 18 Kohlenstoffatomen in der Alk(en)ylkette oder deren Salze einzusetzen. Als weitere anionische Tenside kommen Fettsäure-Derivate von Aminosäuren, beispielsweise von N-Methyltaurin (Tauride) und/oder von N-Methylglycin (Sarkoside) in Betracht. Insbesondere bevorzugt sind dabei die Sarkoside beziehungsweise die Sarkosinate und hier vor allem Sarkosinate von höheren und gegebenenfalls einfach oder mehrfach ungesättigten Fettsäuren wie Oleylsarkosinat. Als weitere anionische Tenside kommen insbesondere Seifen in Betracht. Geeignet sind insbesondere gesättigte Fettsäureseifen, wie die Salze der Laurinsäure, Myristinsäure, Palmitinsäure, Stearinsäure, hydrierten Erucasäure und Behensäure sowie insbesondere aus natürlichen Fettsäuren, zum Beispiel Kokos-, Palmkern- oder Talgfettsäuren, abgeleitete Seifengemische. Zusammen mit diesen Seifen oder als Ersatzmittel für Seifen können auch die bekannten Alkenylbernsteinsäuresalze eingesetzt werden.The polyhydroxy fatty acid amides are preferably derived from reducing sugars having 5 or 6 carbon atoms, in particular from glucose. The group of polyhydroxy fatty acid amides also includes compounds of the formula (IV)

in the R ^{3 is} a linear or branched alkyl or alkenyl radical having 7 to 12 carbon atoms, R ^{4 is} a linear, branched or cyclic alkylene radical or an arylene radical having 2 to 8 carbon atoms and R ^{5 is} a linear, branched or cyclic alkyl radical or a Aryl radical or an oxy-alkyl radical having 1 to 8 carbon atoms, wherein C ₁ -C ₄ alkyl or phenyl radicals are preferred, and [Z] is a linear polyhydroxyalkyl radical whose alkyl chain is substituted with at least two hydroxyl groups, or alkoxylated, preferably ethoxylated or propoxylated derivatives of this group. [Z] is also obtained here preferably by reductive amination of a sugar such as glucose, fructose, maltose, lactose, galactose, mannose or xylose. The N-alkoxy- or N-aryloxy-substituted compounds can then be converted into the desired polyhydroxy fatty acid amides, for example by reaction with fatty acid methyl esters in the presence of an alkoxide as catalyst. Another class of preferred nonionic surfactants used either as the sole nonionic surfactant or in combination with other nonionic surfactants, in particular together with alkoxylated fatty alcohols and / or alkyl glycosides, are alkoxylated, preferably ethoxylated or ethoxylated and propoxylated fatty acid alkyl esters, preferably from 1 to 4 carbon atoms in the alkyl chain, especially fatty acid methyl ester. Nonionic surfactants of the amine oxide type, for example N-cocoalkyl-N, N-dimethylamine oxide and N-tallowalkyl-N, N-dihydroxyethylamine oxide, and the fatty acid alkanolamides may also be suitable. The amount of these nonionic surfactants is preferably not more than that of the ethoxylated fatty alcohols, especially not more than half thereof. Other suitable surfactants are so-called gemini surfactants. These are generally understood as meaning those compounds which have two hydrophilic groups per molecule. These groups are usually separated by a so-called "spacer". This spacer is typically a carbon chain that should be long enough for the hydrophilic groups to be spaced sufficiently apart for them to act independently of each other. Such surfactants are generally characterized by an unusually low critical micelle concentration and the ability to greatly reduce the surface tension of the water. In exceptional cases, the term gemini surfactants not only such "dimer", but also corresponding to "trimeric" surfactants understood. Suitable gemini surfactants are, for example, sulfated hydroxy mixed ethers or dimer alcohol bis and trimer alcohol tris sulfates and ether sulfates. End-capped dimeric and trimeric mixed ethers are characterized in particular by their bi- and multi-functionality. Thus, the end-capped surfactants mentioned have good wetting properties and are low foaming, so that they are particularly suitable for use in machine washing or cleaning processes. However, it is also possible to use gemini-polyhydroxy fatty acid amides or poly-polyhydroxy fatty acid amides. Also suitable are the sulfuric acid monoesters of straight-chain or branched C ₇ -C ₂₁ -alcohols ethoxylated with from 1 to 6 mol of ethylene oxide, such as 2-methyl-branched C ₉ -C ₁₁ -alcohols having on average 3.5 mol of ethylene oxide (EO) or C ₁₂ - C ₁₈ -fatty alcohols with 1 to 4 EO. The preferred anionic surfactants also include the salts of alkylsulfosuccinic acid, which are also referred to as sulfosuccinates or as sulfosuccinic acid esters, and the monoesters and / or diesters of sulfosuccinic acid with alcohols, preferably fatty alcohols and in particular ethoxylated fatty alcohols. Preferred sulfosuccinates contain C ₈ to C ₁₈ fatty alcohol residues or mixtures of these. Particularly preferred sulfosuccinates contain a fatty alcohol residue derived from ethoxylated fatty alcohols, which by themselves are nonionic surfactants. Sulfosuccinates, whose fatty alcohol residues are derived from ethoxylated fatty alcohols with a narrow homolog distribution, are again particularly preferred. Likewise, it is also possible to use alk (en) ylsuccinic acid having preferably 8 to 18 carbon atoms in the alk (en) yl chain or salts thereof. Suitable further anionic surfactants are fatty acid derivatives of amino acids, for example N-methyltaurine (Tauride) and / or N-methylglycine (sarcosides). Particularly preferred are the sarcosides or the sarcosinates and here especially sarcosinates of higher and optionally monounsaturated or polyunsaturated fatty acids such as oleyl sarcosinate. As further anionic surfactants are particularly soaps into consideration. Particularly suitable are saturated fatty acid soaps, such as the salts of lauric acid, myristic acid, palmitic acid, stearic acid, hydrogenated erucic acid and behenic acid and, in particular, soap mixtures derived from natural fatty acids, for example coconut, palm kernel or tallow fatty acids. Together with these soaps or as a substitute for soaps, it is also possible to use the known alkenylsuccinic acid salts.

The anionic surfactants, including soaps, may be in the form of their sodium, potassium or ammonium salts and as soluble salts of organic bases, such as mono-, di- or triethanolamine. In front Preferably, the anionic surfactants are in the form of their sodium or potassium salts, especially in the form of the sodium salts.

surfactants are in proportions in inventive compositions of preferably 5% by weight to 50% by weight, in particular 8% by weight to 30 wt .-%, contained.

An agent according to the invention preferably contains at least one water-soluble and / or water-insoluble, organic and / or inorganic builder. The water-soluble organic builder substances include polycarboxylic acids, in particular citric acid and sugar acids, monomeric and polymeric aminopolycarboxylic acids, in particular methylglycinediacetic acid, nitrilotriacetic acid and ethylenediaminetetraacetic acid and polyaspartic acid, polyphosphonic acids, in particular aminotris (methylenephosphonic acid), ethylenediaminetetrakis (methylenephosphonic acid) and 1-hydroxyethane-1,1-diphosphonic acid, polymeric hydroxy compounds such as dextrin and also polymeric (poly) carboxylic acids, in particular the polycarboxylates obtainable by oxidation of polysaccharides or dextrins, polymeric acrylic acids, methacrylic acids, maleic acids and copolymers thereof, which may also contain polymerized small amounts of polymerizable substances without carboxylic acid functionality. The molecular weight of the homopolymers of unsaturated carboxylic acids is generally between 3,000 and 200,000, of the copolymers between 2,000 and 200,000, preferably 30,000 to 120,000, each based on the free acid. A particularly preferred acrylic acid-maleic acid copolymer has a molecular weight of from 30,000 to 100,000. Commercial products are, for example Sokalan ^® CP 5, CP 10 and PA 30 from BASF. Suitable, although less preferred, compounds of this class are copolymers of acrylic or methacrylic acid with vinyl ethers, such as vinylmethyl ethers, vinyl esters, ethylene, propylene and styrene, in which the acid content is at least 50% by weight. It is also possible to use terpolymers which contain two unsaturated acids and / or salts thereof as monomers and also vinyl alcohol and / or an esterified vinyl alcohol or a carbohydrate as the third monomer as water-soluble organic builder substances. The first acidic monomer or its salt is derived from a monoethylenically unsaturated C ₃ -C ₈ -carboxylic acid and preferably from a C ₃ -C ₄ -monocarboxylic acid, in particular from (meth) -acrylic acid. The second acidic monomer or its salt may be a derivative of a C ₄ -C ₈ -dicarboxylic acid, with maleic acid being particularly preferred, and / or a derivative of an allylsulfonic acid which is substituted in the 2-position by an alkyl or aryl radical. Such polymers generally have a molecular weight between 1,000 and 200,000. Further preferred copolymers are those which preferably have as monomers acrolein and acrylic acid / acrylic acid salts or vinyl acetate. The organic builder substances can be used, in particular for the preparation of liquid agents, in the form of aqueous solutions, preferably in the form of 30 to 50 percent by weight aqueous solutions. All of the acids mentioned are generally used in the form of their water-soluble salts, in particular their alkali metal salts.

such If desired, organic builders may be used in amounts up to 40 wt .-%, in particular up to 25 wt .-% and preferably from 1% by weight to 8% by weight. Quantities near the mentioned Upper limit are preferably in pasty or liquid, in particular hydrous, inventive Funds used.

Suitable water-soluble inorganic builder materials are, in particular, alkali metal silicates, alkali metal carbonates and alkali metal phosphates, which may be in the form of their alkaline, neutral or acidic sodium or potassium salts. Examples of these are trisodium phosphate, tetrasodium diphosphate, disodium dihydrogen diphosphate, pentasodium triphosphate, so-called sodium hexametaphosphate, oligomeric trisodium phosphate with degrees of oligomerization of 5 to 1000, in particular 5 to 50, and the corresponding potassium salts or mixtures of sodium and potassium salts. Crystalline or amorphous alkali metal aluminosilicates, in amounts of up to 50% by weight, preferably not more than 40% by weight, and in liquid agents, in particular from 1% by weight to 5% by weight, are particularly suitable as water-insoluble, water-dispersible inorganic builder materials. used. Among these, the crystalline sodium aluminosilicates in detergent quality, more particularly zeolite A, P and optionally X, alone or in mixtures, for example in the form of a co-crystals of zeolites A and X (VEGOBOND ^® AX, a commercial product of Condea August SpA), preferably , Amounts near the above upper limit are preferably used in solid, particulate agents. In particular, suitable aluminosilicates have no particles with a particle size greater than 30 .mu.m and preferably consist of at least 80% by weight of particles having a size of less than 10 .mu.m. Their calcium binding capacity, according to the information of the German Patent DE 24 12 837 can be determined, is usually in the range of 100 to 200 mg CaO per gram.

Suitable substitutes or partial substitutes for the said aluminosilicate are crystalline alkali silicates which may be present alone or in a mixture with amorphous silicates. The in the invention The alkali metal silicates which are useful as builders preferably have a molar ratio of alkali metal oxide to SiO _{2 of} less than 0.95, in particular of from 1: 1.1 to 1:12, and may be present in amorphous or crystalline form. Preferred alkali metal silicates are the sodium silicates, in particular the amorphous sodium silicates, with a molar ratio of Na ₂ O: SiO ₂ of 1: 2 to 1: 2.8. The crystalline silicates which may be present alone or in admixture with amorphous silicates, are crystalline layer silicates with the general formula Na ₂ Si _x O _{2x + 1} · are used yH ₂ O, in which x, the so-called module, a number from 1.9 to 22, in particular 1.9 to 4 and y is a number from 0 to 33 and preferred values for x are 2, 3 or 4. Preferred crystalline phyllosilicates are those in which x in the abovementioned general formula assumes the values 2 or 3. In particular, both β- and δ-sodium disilicates (Na ₂ Si ₂ O ₅ .yH ₂ O) are preferred. Also prepared from amorphous alkali silicates, practically anhydrous crystalline alkali silicates of the abovementioned general formula in which x is a number from 1.9 to 2.1, can be used in inventive compositions. In a further preferred embodiment of the composition according to the invention, a crystalline sodium layer silicate with a modulus of 2 to 3 is used, as can be prepared from sand and soda. Crystalline sodium silicates with a modulus in the range from 1.9 to 3.5 are used in a further preferred embodiment of compositions according to the invention. Crystalline layer-form silicates of formula (I) given above are sold by Clariant GmbH under the trade name Na-SKS, eg. Na-SKS-1 (Na ₂ Si ₂₂ O ₄₅ .xH ₂ O, Kenyaite), Na-SKS-2 (Na ₂ Si ₁₄ O ₂₉ .xH ₂ O, magadiite), Na-SKS-3 (Na ₂ Si ₈ O ₁₇ .xH ₂ O) or Na-SKS-4 (Na ₂ Si ₄ O ₉ .xH ₂ O, Makatite). Of these, especially Na-SKS-5 (α-Na ₂ Si ₂ O ₅ ), Na-SKS-7 (β-Na ₂ Si ₂ O ₅ , natrosilite), Na-SKS-9 (NaHSi ₂ O ₅ · 3H ₂ O), Na-SKS-10 (NaHSi ₂ O ₅ · 3H ₂ O, kanemite), Na-SKS-11 (t-Na ₂ Si ₂ O ₅₎ and Na-SKS-13 (NaHSi ₂ O ₅ ), but especially Na-SKS-6 (δ-Na ₂ Si ₂ O ₅ ). In a preferred embodiment of inventive compositions is a granular compound of crystalline phyllosilicate and citrate, of crystalline layered silicate and aforementioned (co) polymeric polycarboxylic acid or of alkali silicate and alkali metal as it is available, for example under the name Nabion ^® 15 commercially ,

builders are preferred in the compositions according to the invention in amounts up to 75 wt .-%, in particular 5 wt .-% to 50 included.

When for use in the invention Means of suitable peroxygen compounds come in particular organic peracids or peracid salts organic Acids, such as phthalimidopercaproic acid, perbenzoic acid or salts of diperdodecanedioic acid, hydrogen peroxide and under the washing conditions hydrogen peroxide-releasing inorganic Salts, which include perborate, percarbonate, persilicate and / or persulfate like Caroat belong. If solid peroxygen compounds can be used, they can be in the form of powders or granules are used, which are also known in principle Wrapped up. If an inventive Agent contains peroxygen compounds, these are in Amounts of preferably up to 50% by weight, in particular of 5% by weight to 30 wt .-%, present. The addition of small amounts of known bleach stabilizers such as phosphonates, borates or metaborates and metasilicates and magnesium salts such as magnesium sulfate be expedient.

When Bleach activators may be compounds that are under perhydrolysis conditions aliphatic peroxycarboxylic acids with preferably 1 to 10 C-atoms, in particular 2 to 4 C-atoms, and / or optionally substituted Perbenzoic acid, are used. Are suitable Substances which contain O- and / or N-acyl groups of said C atomic number and / or optionally substituted benzoyl groups. Preferred are multiply acylated alkylenediamines, in particular tetraacetylethylenediamine (TAED), acylated triazine derivatives, in particular 1,5-diacetyl-2,4-dioxohexahydro-1,3,5-triazine (DADHT), acylated glycolurils, especially tetraacetylglycoluril (TAGU), N-acylimides, especially N-nonanoylsuccinimide (NOSI), acylated Phenolsulfonates, especially n-nonanoyl or isononanoyloxybenzenesulfonate (n- or iso-NOBS), carboxylic anhydrides, in particular Phthalic anhydride, acylated polyhydric alcohols, in particular Triacetin, ethylene glycol diacetate, 2,5-diacetoxy-2,5-dihydrofuran and enol ester, as well as acetylated sorbitol and mannitol, respectively their described mixtures (SORMAN), acylated sugar derivatives, in particular pentaacetylglucose (PAG), pentaacetyl-fructose, tetraacetylxylose and octaacetyl lactose and acetylated, optionally N-alkylated Glucamine and gluconolactone, and / or N-acylated lactams, for example N-benzoyl. The hydrophilic substituted acylacetals and The acyl lactams are also preferably used. Also combinations Conventional bleach activators can be used. Such bleach activators can, especially in the presence above hydrogen peroxide-providing bleach, in the usual Amount range, preferably in amounts of 0.5 wt .-% to 10 wt .-%, in particular from 1% by weight to 8% by weight, based on the total agent, be missing when using percarboxylic acid as sole bleach, however, preferably completely.

In addition to, or in place of, the conventional bleach activators, sulfonimines may also be used and / or bleach-enhancing transition metal salts or transition metal complexes may be present as so-called bleach catalysts.

To in the inventive compositions, in particular if they are in liquid or pasty form, include water-usable organic solvents Alcohols having 1 to 4 carbon atoms, in particular methanol, ethanol, isopropanol and tert-butanol, diols having 2 to 4 C atoms, in particular ethylene glycol and propylene glycol, as well as mixtures thereof and those mentioned Classes of derivable ether. Such water-miscible Solvents are in the inventive Preferably in amounts not exceeding 30% by weight, in particular from 6% to 20% by weight.

to Setting a desired, through the mixture the remaining components are not self-generating pH the agents according to the invention can be and environmentally friendly acids, in particular citric acid, Acetic acid, tartaric acid, malic acid, Lactic acid, glycolic acid, succinic acid, glutaric acid and / or adipic acid, but also mineral acids, especially sulfuric acid, or bases, in particular ammonium or alkali hydroxides. Such pH regulators are in the inventive compositions in amounts of preferably not more than 20% by weight, in particular from 1.2% by weight to 17% by weight, contain.

graying have the task of removing the dirt from the textile fiber to keep suspended in the fleet. These are water-soluble Colloids mostly organic nature suitable, such as starch, Glue, gelatin, salts of ether carboxylic acids or ether sulfonic acids starch or cellulose or salts of acidic sulfuric acid esters cellulose or starch. Also water-soluble, acidic Group-containing polyamides are suitable for this purpose. Furthermore, other than the above-mentioned starch derivatives can be used use, for example, aldehyde levels. To be favoured Cellulose ethers, such as carboxymethyl cellulose (Na salt), methyl cellulose, Hydroxyalkylcellulose and mixed ethers, such as methylhydroxyethylcellulose, Methylhydroxypropylcellulose, methylcarboxymethylcellulose and their Mixtures, for example in amounts of 0.1 to 5 wt .-%, based on the means used.

invention Textile detergents can be used as optical brighteners, for example Derivatives of Diaminostilbendisulfonsäure or their alkali metal salts, although they are for the Use as a color detergent preferably free of optical brighteners are. For example, salts of 4,4'-bis (2-anilino-4-morpholino-1,3,5-triazinyl-6-amino) stilbene-2,2'-disulfonic acid are suitable or similar compounds, instead of the morpholino group a diethanolamino group, a methylamino group, an anilino group or carry a 2-methoxyethylamino group. Furthermore you can Brighteners of the type of substituted diphenylstyrene may be present for example, the alkali salts of 4,4'-bis (2-sulfostyryl) -diphenyl, 4,4'-bis (4-chloro-3-sulfostyryl) -diphenyls, or 4- (4-chlorostyryl) -4 '- (2-sulfostyryl) -diphenyls. Mixtures of the aforementioned optical brightener can be used.

In particular, when used in mechanical processes, it may be advantageous to add conventional foam inhibitors to the compositions. As foam inhibitors are, for example, soaps of natural or synthetic origin, which have a high proportion of C ₁₈ -C ₂₄ fatty acids. Suitable non-surfactant foam inhibitors are, for example, organopolysiloxanes and mixtures thereof with microfine, optionally silanized silica and paraffins, waxes, microcrystalline waxes and mixtures thereof with silanated silicic acid or bis-fatty acid alkylenediamides. It is also advantageous to use mixtures of various foam inhibitors, for example those of silicones, paraffins or waxes. The foam inhibitors, in particular silicone- and / or paraffin-containing foam inhibitors, are preferably bound to a granular, water-soluble or dispersible carrier substance. In particular, mixtures of paraffins and bistearylethylenediamide are preferred.

The to select ingredients as well as the conditions under which the agent is used, such as temperature, pH, ionic strength, redox ratios or mechanical influences, should be for the respective Cleaning problem to be optimized. So are usual temperatures for detergents and cleaners in the range of 10 ° C with manual means over 40 ° C and 60 ° C up to 95 ° for mechanical means or technical Applications. As in modern washing and dishwasher the Temperature is usually infinitely adjustable, are also all intermediate stages the temperature included. Preferably, the ingredients become of the funds concerned. Synergies are preferred in terms of cleaning performance. Especially preferred in this regard are synergies that are in a temperature range between 20 ° C and 60 ° C are present, as well as in the inventive Agents contained in this temperature range catalytically is active.

The Preparation of inventive solid means provides no difficulties and can be done in a known way, for example by spray-drying or granulation, enzymes being used and any other thermally sensitive ingredients like for example bleach, if appropriate later separately be added. For the production according to the invention Agents with increased bulk density, in particular in the range of 650 g / l to 950 g / l, is an extrusion step preferred method.

to Preparation of agents according to the invention in tablet form, the one-phase or multi-phase, monochrome or multicolor and in particular from one or more layers, in particular two layers can exist, it is preferable to proceed in such a way that all components - if necessary one layer each - in a mixer mixed together and the mixture by means of conventional Tablet presses, for example eccentric presses or rotary presses, with compressive forces in the range of about 50 to 100 kN, preferably compressed at 60 to 70 kN. Especially for multi-layered tablets, it may be advantageous if at least a layer is pre-pressed. This is preferably included Pressing forces between 5 and 20 kN, in particular carried out at 10 to 15 kN. You get so easily unbreakable and yet sufficiently fast under conditions of use soluble tablets with breaking and bending strengths of normally 100 to 200 N, but preferably over 150 N. Preferably, a tablet produced in this way has a weight from 10 g to 50 g, especially from 15 g to 40 g. The spatial form the tablets are arbitrary and can be round, oval or angular, although intermediate forms are possible. Corners and edges are advantageously rounded. Round tablets preferably have a diameter of 30 mm to 40 mm. Especially the size of square or cuboid tablets, which predominantly over the metering device, for example, the dishwasher is introduced depends on the geometry and the volume of this metering device. Exemplary preferred embodiments have a footprint of (20 to 30 mm) × (34 to 40 mm), in particular 26 × 36 mm or 24 × 38 mm up.

liquid or pasty invention Agent in the form of common solvents Solutions are usually made by simply mixing the Ingredients that are in substance or as a solution in one automatic mixer can be given.

embodiments The present invention thus includes all such fixed, powdery, liquid, gelatinous or pasty dosage forms of the agents, if appropriate can also consist of several phases as well as in compressed or uncompressed form. Another Embodiment of the invention therefore represent means which are characterized in that they are as one-component system available. Such means preferably consist of one phase. Of course However, agents according to the invention can also consist of several phases. In a further embodiment of the Invention, the detergent or cleaning agent is therefore characterized are that it is divided into several components.

To the solid dosage forms according to the invention include extrudates, granules, tablets or pouches, which are packed both in large containers and in portions may be present. Alternatively, the agent is as free-flowing Powder, in particular with a bulk density of 300 g / l to 1200 g / l, in particular 500 g / l to 900 g / l or 600 g / l up to 850 g / l.

Further agents according to the invention can also be liquid, be gelatinous or pasty. Another embodiment The invention is therefore characterized in that the washing or detergent in liquid, gel or pasty form, especially in the form of a non-aqueous liquid detergent or a non-aqueous paste or in the form of a aqueous liquid detergent or a hydrous paste.

The Detergents or cleaning agents according to the invention can in a container, preferably an air-permeable Container, be packaged, out of it just before use or released during the washing process. Especially may further contain the protease contained in the agent and / or further Ingredients of the agent with a room temperature or at Absence of water for the enzyme impermeable substance which under application conditions of the agent becomes permeable to the enzyme. Such an embodiment The invention is thus characterized in that the protease with a at room temperature or in the absence of water for the protease impermeable substance envelops is.

Compositions according to the invention may contain only one protease as described. Alternatively, however, they may contain other proteases or other enzymes in a concentration effective for the effectiveness of the agent. A further subject of the invention thus represents a washing or cleaning agent, which is characterized in that it comprises at least one further enzyme, esp in particular a protease, amylase, cellulase, hemicellulase, mannanase, taunase, xylanase, xanthanase, β-glucosidase, carrageenase, oxidase, oxidoreductase, lipase or combinations thereof, wherein in principle all known in the art for this purpose enzymes can be used. The enzymes may be adsorbed to carriers and / or embedded in encapsulants to protect against premature inactivation. They are in the washing or cleaning agents according to the invention preferably in amounts of 1 × 10 ^-8 to 5 percent by weight based on active protein. Preferably, the enzymes are from 0.001 to 5% by weight, more preferably from 0.01 to 5% by weight, even more preferably from 0.05 to 4% by weight and most preferably from 0.075 to 3.5% by weight. -%, wherein each enzyme contained may be present in the said amount. If several enzymes are to be used in the agent according to the invention, this can be carried out by incorporation of the two or more separate or in a known manner separately prepared enzymes or by two or more enzymes formulated together in a granule.

The protein concentration can be determined by known methods, for example the BCA method (bicinchoninic acid, 2,2'-biquinolyl-4,4'-dicarboxylic acid) or the biuret method ( Gornall AG, CS Bardawill and MM David, J. Biol. Chem., 177 (1948), pp. 751-766 ). The further enzymes preferably support the effect of the agent, for example the cleaning performance of a washing or cleaning agent, with regard to certain stains or stains. Particularly preferably, the enzymes show synergistic effects with respect to their action against certain stains or stains, ie the enzymes contained in the middle composition mutually support each other in their cleaning performance. Very particular preference is given to such synergism between the protease from a bacterium of the genus Xanthomonas and another enzyme of an agent according to the invention, including in particular between said protease and an amylase and / or a mannanase and / or a lipase. Synergistic effects can occur not only between different enzymes, but also between one or more enzymes and other ingredients of the composition according to the invention.

Among the proteases, those of the subtilisin type are preferable. Examples thereof are the subtilisins BPN 'and Carlsberg, the protease PB92, the subtilisins 147 and 309, the alkaline protease from Bacillus lentus, subtilisin DY and the enzymes thermitase, proteinase K and the subtilases, but not the subtilisins in the narrower sense Proteases TW3 and TW7. Subtilisin Carlsberg in a developed form under the trade names Alcalase ^® from Novozymes A / S, Bagsvaerd, Denmark. The subtilisins 147 and 309 are sold under the trade names Esperase ^®, or Savinase ^® from Novozymes. From the protease from Bacillus lentus DSM 5483 derived under the name BLAP ^® protease variants derived. Other usable proteases are, for example, under the trade names Durazym ^®, relase ^®, Everlase® ^®, Nafizym, Natalase ^®, Kannase® ^® and Ovozymes ^® from Novozymes, under the trade names Purafect ^®, Purafect ^® OxP and Properase.RTM ^® by the company Genencor, that under the trade name Protosol® ^® from Advanced Biochemicals Ltd., Thane, India, under the trade name Wuxi ^® from Wuxi Snyder Bioproducts Ltd., China, under the trade names Proleather® ^® and protease P ^® by the company Amano Pharmaceuticals Ltd., Nagoya, Japan, and the enzyme available under the name Proteinase K-16 from Kao Corp., Tokyo, Japan.

The protease activity in agents according to the invention can be determined according to the method described in Surfactants, Vol. 7 (1970), pp. 125-132 , described method can be determined. It is accordingly stated in PE (protease units). A composition according to the invention preferably has a proteolytic activity in the range of about 100 PE / g to about 10,000 PE / g, in particular 300 PE / g to 8000 PE / g.

Examples of amylases which can be synthesized according to the invention are the α-amylases from Bacillus licheniformis, from Bacillus amyloliquefaciens or from Bacillus stearothermophilus and also their further developments improved for use in detergents or cleaners. The enzyme from Bacillus licheniformis is available from Novozymes under the name Termamyl ^® and from Genencor under the name Purastar® ^® ST. Development products of this α-amylase are available from Novozymes under the trade names Duramyl ^® and Termamyl ^® ultra, from Genencor under the name Purastar® ^® OxAm and from Daiwa Seiko Inc., Tokyo, Japan, as Keistase ^®. The α-amylase from Bacillus amyloliquefaciens is sold by Novozymes under the name BAN ^®, and variants derived from the α-amylase from Bacillus stearothermophilus BSG under the name ^® and Novamyl ^®, also from Novozymes.

Furthermore, for this purpose, the α-amylase from Bacillus sp. A 7-7 (DSM 12368) and the cyclodextrin glucanotransferase (CGTase) from B. agaradherens (DSM 9948). Furthermore, the amylolytic enzymes belonging to the sequence space of α-amylases can be used, which in the international Patent application WO 03/002711 A2 is defined, and the ones in the application WO 03/054177 A2 to be discribed. Likewise, fusion products of said molecules can be used.

In addition, the enhancements available under the trade names Fungamyl.RTM ^® by Novozymes of α-amylase from Aspergillus niger and A. oryzae, which are. Further usable commercial products are, for example, the amylase LT ^® and Stainzyme ^® or Stainzyme ultra ^® or Stainzyme plus ^® , the latter also from the company Novozymes. Variants of these enzymes which can be obtained by point mutations can also be incorporated into agents according to the invention.

Compositions according to the invention may contain lipases or cutinases, in particular because of their triglyceride-cleaving activities, but also in order to generate in situ peracids from suitable precursors. These include, for example, the lipases originally obtainable from Humicola lanuginosa (Thermomyces lanuginosus) or further developed, in particular those with the amino acid exchange D96L. They are sold, for example, by Novozymes under the trade names Lipolase ^®, Lipolase Ultra ^®, LipoPrime® ^®, Lipozyme® ^® and Lipex ^®. Furthermore, for example, the cutinases can be used, which were originally isolated from Fusarium solani pisi and Humicola insolens. Likewise useable lipases are available from Amano under the designations Lipase CE ^®, Lipase P ^®, Lipase B ^®, or lipase CES ^®, Lipase AKG ^®, Bacillis sp. ^Lipase® , Lipase ^AP® , Lipase M- ^AP® and Lipase ^{AML® are} available. From the company Genencor, for example, the lipases, or cutinases can be used, the initial enzymes were originally isolated from Pseudomonas mendocina and Fusarium solanii. Other important commercial products are the originally marketed by Gist-Brocades preparations M1 Lipase ^® and Lipomax® ^® and the enzymes marketed by Meito Sangyo KK, Japan under the names Lipase MY-30 ^®, Lipase OF ^® and lipase PL ^® to mention also the product Lumafast® ^® from Genencor.

invention Means can, especially if they are for treatment of textiles are thought to contain cellulases, depending on the purpose as pure enzymes, as enzyme preparations or in the form of Mixtures in which the individual components advantageously with regard to their different performance aspects. These performance aspects include, in particular, contributions to the primary washing power, to the secondary washing power of the agent (anti-redeposition effect or graying inhibition) and conditioning (fabric effect), to the exercise of a "stone wash "effect.

A usable fungal, Endoglucanase (EG) -rich cellulase preparation, or their further developments are offered by the company Novozymes under the trade name Celluzyme ^® . The products Endolase® ^® and Carezyme ^®, likewise available from Novozymes, are based on the 50 kD EG and 43 kD EG from H. insolens DSM 1800. Further commercial products of this company are Cellusoft® ^® and Renozyme ^®. Also usable are for example the 20 kD EG Melanocarpus, which is available from AB Enzymes, Finland, under the trade names Ecostone ^® and Biotouch ^®. Further commercial products from AB Enzymes are Econase® ^® and ECOPULP ^®. Other suitable cellulases are from Bacillus sp. CBS 670.93 and CBS 669.93, those derived from Bacillus sp. CBS 670.93 from the company Genencor under the trade name Puradax ^{® is} available. Further commercial products of the company Genencor are "Genencor detergent cellulase L" and IndiAge ^® Neutra.

Compositions according to the invention may contain, in particular for the removal of certain problem soiling, further enzymes which are combined under the term hemicellulases. These include, for example, mannanases, xanthan lyases, pectin lyases (= pectinases), pectin esterases, pectate lyases, xyloglucanases (= xylanases), pullulanases and β-glucanases. In this regard, suitable enzymes United States for example, under the name Gamanase ^® and Pektinex AR ^® from Novozymes, under the name Rohapec ^® B1 from AB Enzymes and under the name Pyrolase® ^® from Diversa Corp., San Diego, CA. , The recovered from Bacillus subtilis β-glucanase is available under the name Cereflo ^® from Novozymes. According to the invention particularly preferred hemicellulases are mannanases, which are marketed under the trade names man Away ^® by the company Novozymes or Purabrite ^® by the company Genencor.

To increase the bleaching effect agents of the invention may also oxidoreductases, such as oxidases, oxygenases, catalases (which react at low H ₂ O ₂ concentrations as peroxidase), peroxidases, such as halo-, chloro-, bromo-, lignin, glucose or Manganese peroxidases, dioxygenases or laccases (phenol oxidases, polyphenol oxidases). Suitable commercial products Denilite® ^® 1 and 2 from Novozymes should be mentioned. As an example of advantageous systems for enzymatic perhydrolysis can be applied to the applications WO 98/45398 A1 . WO 2005/056782 A2 such as WO 2004/058961 A1 directed. A combined enzymatic bleaching system comprising an oxidase and a perhydrolase describes the application WO 2005/124012 , Advantageously, it is additionally preferred to add organic, particularly preferably aromatic, compounds which interact with the enzymes in order to enhance the activity of the relevant oxidoreductases (enhancers) or to ensure the flow of electrons (mediators) at greatly varying redox potentials between the oxidizing enzymes and the soils.

at The comparison of the performance of two detergent enzymes must be between distinguished from the same and the same activity become. Especially when genetically engineered, largely free of side activity Enzyme preparations are appropriate for the same protein use. Because that is a statement about whether it is possible the same amount of protein - as a measure of the yield of fermentative production - comparable Results. Gap the respective conditions from active substance to total protein (the values of the specific activity) apart, so is an activity-same comparison too recommend because this is the respective enzymatic properties be compared. Generally speaking, a low specific Activity by adding a larger one Protein amount can be compensated. This is ultimately for an economic consideration.

Table 1 below shows formulations for detergents and cleaners according to the invention. Each formulation in this case comprises a protease from the bacterium mentioned and at least the particular detergent ingredient specified in the stated range of amounts. Each recipe may optionally include other ingredients. Table 1: Protease out Builder substance 0.1-75% by weight Surfactant surfactant 5-50% by weight Peroxygen compound as a bleaching agent 0.1-50% by weight Bleach activator 0.5-10% by weight Xanthomonas albilineans Recipe 1 Recipe 2 Recipe 3 Recipe 4 Xanthomonas alfalfae Recipe 5 Recipe 6 Recipe 7 Recipe 8 Xanthomonas alfalfae subsp. alfalfae Recipe 9 Recipe 10 Recipe 11 Recipe 12 Xanthomonas alfalfae subsp. citrumelonis Recipe 13 Recipe 14 Recipe 15 Recipe 16 Xanthomonas ampelina (Xylophilus ampelinus) Recipe 17 Recipe 18 Recipe 19 Recipe 20 Xanthomonas arboricola Recipe 21 Recipe 22 Recipe 23 Recipe 24 Xanthomonas axonopodis Recipe 25 Recipe 26 Recipe 27 Recipe 28 Xanthomonas bromi Recipe 29 Recipe 30 Recipe 31 Recipe 32 Xanthomonas campestris Recipe 33 Recipe 34 Recipe 35 Recipe 36 Xanthomonas cassavae Recipe 37 Recipe 38 Recipe 39 Recipe 40 Xanthomonas citri Recipe 41 Recipe 42 Recipe 43 Recipe 44 Xanthomonas citri subsp. citri Recipe 45 Recipe 46 Recipe 47 Recipe 48 Xanthomonas citri subsp. malvacearum Recipe 49 Recipe 50 Recipe 51 Recipe 52 Xanthomonas codiaei Recipe 53 Recipe 54 Recipe 55 Recipe 56 Xanthomonas cucurbitae Recipe 57 Recipe 58 Recipe 59 Recipe 60 Xanthomonas cynarae Recipe 61 Recipe 62 Recipe 63 Recipe 64 Xanthomonas euvesicatoria Recipe 65 Recipe 66 Recipe 67 Recipe 68 Xanthomonas fragariae Recipe 69 Recipe 70 Recipe 71 Recipe 72 Xanthomonas fuscans Recipe 73 Recipe 74 Recipe 75 Recipe 76 Xanthomonas fuscans subsp. aurantifolii Recipe 77 Recipe 78 Recipe 79 Recipe 80 Xanthomonas fuscans subsp. fuscans Recipe 81 Recipe 82 Recipe 83 Recipe 84 Xanthomonas gardneri Recipe 85 Recipe 86 Recipe 87 Recipe 88 Xanthomonas hortorum Recipe 89 Recipe 90 Recipe 91 Recipe 92 Xanthomonas hyacinthi Recipe 93 Recipe 94 Recipe 95 Recipe 96 Xanthomonas melonis Recipe 97 Recipe 98 Formulation 99 Recipe 100 Xanthomonas oryzae Recipe 101 Recipe 102 Recipe 103 Recipe 104 Xanthomonas perforans Recipe 105 Recipe 106 Recipe 107 Recipe 108 Xanthomonas phaseoli Recipe 109 Recipe 110 Recipe 111 Recipe 112 Xanthomonas pisi Recipe 113 Recipe 114 Recipe 115 Recipe 116 Xanthomonas populi Recipe 117 Recipe 118 Recipe 119 Recipe 120 Xanthomonas sacchari Recipe 121 Recipe 122 Recipe 123 Recipe 124 Xanthomonas theicola Recipe 125 Recipe 126 Recipe 127 Recipe 128 Xanthomonas translucens Recipe 129 Recipe 130 Recipe 131 Recipe 132 Xanthomonas vasicola Recipe 133 Recipe 134 Recipe 135 Recipe 136 Xanthomonas vesicatoria Recipe 137 Recipe 138 Recipe 139 Recipe 140 Xanthomonas axonopodis DSM 3585 Recipe 141 Recipe 142 Recipe 143 Recipe 144 Xanthomonas axonopodis pvar. Begoniae DSM 50850 Recipe 145 Recipe 146 Recipe 147 Recipe 148 Xanthomonas axonopodis pvar. malvacearum DSM 1220 Recipe 149 Recipe 150 Recipe 151 Recipe 152 Xanthomonas campestris DSM 1050 (NCPPB 1929) Recipe 153 Recipe 154 Recipe 155 Recipe 156 Xanthomonas campestris DSM 19000 Recipe 157 Recipe 158 Recipe 159 Recipe 160 Xanthomonas campestris pvar. campestris DSM 3586T Recipe 161 Recipe 162 Recipe 163 Recipe 164 Xanthomonas campestris pvar. pelargonii DSM 50857 Recipe 165 Recipe 166 Recipe 167 Recipe 168

All formulations shown in Table 1 may include other detergent ingredients. In the following Table 2, therefore, the formulations of Table 1 are given as basic formulations with an additionally present in the respective base formulation detergent ingredient in the specified amount. Here, each base formulation with each ingredient represents a separate, independent recipe. For example, the recipe 1 with an organic solvent, with a pH regulator, with an optical brightener and with a grayness inhibitor as four independent egg recipes to understand. Table 2: Recipe no. Organic solvent pH regulator Optical brightener antiredeposition 1 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 2 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 3 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 4 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 5 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 6 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 7 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 8th 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 9 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 10 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 11 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 12 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 13 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 14 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 15 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 16 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 17 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 18 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 19 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 20 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 21 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 22 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 23 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 24 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 25 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 26 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 27 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 28 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 29 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 30 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 31 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 32 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 33 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 34 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 35 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 36 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 37 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 38 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 39 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 40 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 41 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 42 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 43 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 44 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 45 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 46 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 47 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 48 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 49 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 50 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 51 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 52 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 53 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 54 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 55 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 56 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 57 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 58 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 59 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 60 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 61 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 62 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 63 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 64 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 65 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 66 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 67 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 68 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 69 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 70 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 71 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 72 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 73 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 74 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 75 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 76 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 77 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 78 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 79 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 80 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 81 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 82 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 83 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 84 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 85 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 86 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 87 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 88 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 89 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 90 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 91 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 92 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 93 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 94 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 95 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 96 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 97 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 98 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 99 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 100 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 101 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 102 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 103 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 104 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 105 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 106 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 107 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 108 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 109 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 110 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 111 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 112 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 113 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 114 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 115 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 116 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 117 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 118 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 119 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 120 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 121 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 122 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 123 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 124 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 125 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 126 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 127 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 128 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 129 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 130 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 131 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 132 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 133 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 134 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 135 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 136 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 137 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 138 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 139 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 140 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 141 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 142 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 143 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 144 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 145 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 146 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 147 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 148 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 149 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 150 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 151 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 152 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 153 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 154 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 155 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 156 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 157 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 158 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 159 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 160 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 161 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 162 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 163 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 164 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 165 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 166 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 167 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight 168 0.1-30% by weight 0.1-20% by weight 0.1-5% by weight 0.1-5% by weight

a own subject of the invention provides the use of an inventive Detergent or cleaning agent for the removal of protease-sensitive Soiling on textiles or hard surfaces, d. H. for cleaning textiles or hard surfaces, represents.

Because Detergents or cleaning agents according to the invention can be used to remove from textiles or from hard surfaces to eliminate proteinaceous contaminants or at least reduce it. Provide embodiments for example, hand washing, manual removal of patches of textiles or of hard surfaces or the use in connection with a machine process represents.

In a preferred embodiment of this use the relevant inventive washing or Cleaning agent according to one of the embodiments described above provided.

a another own subject of the invention provide methods for cleaning textiles or hard surfaces in which at least in one of the method steps an inventive Medium is used. The procedure for cleaning textiles or hard surfaces is thus characterized that in at least one method step, an inventive Medium is applied.

this includes fall both manual and mechanical procedures, whereby machine Method because of their more precise controllability, what For example, the amounts used and exposure times, preferably are.

Such Methods are generally characterized by the fact that in several Process steps to various cleaning-active substances applied the cleaning material and washed off after the contact time or that the items to be cleaned in any other way with a Detergent or a solution or dilution this agent - the so-called wash liquor - treated becomes. All conceivable washing or cleaning methods can in at least one of the method steps to the application of a enriched agent according to the invention and then provide embodiments of the present invention represents.

As already explained above, it is a further object of the present invention to provide protease enzymes which are suitable for use in agents according to the invention and are characterized by one characterized by a good, preferably by an improved proteolytic activity with respect to at least one soiling, preferably with respect to several soils, in the above-mentioned temperature range when used in agents according to the invention.

Solved This task is characterized by a protease that is characterized is that they are naturally occurring in a bacterium of the genus Xanthomonas is present and they are in an aqueous solution in the presence of at least one surfactant, a proteolysis index of at least 1.1. The proteolysis index is increasingly preferred at least 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.25, 3, 5, 3.75, 4.0, 4.25, 4.5, 4.75 and 5.0.

This Proteolysis index is determined using standardized polluters Textiles issued by the Federal Material Testing and testing institute, St. Gallen, Switzerland (EMPA), or the laundry research institute, Krefeld, can be obtained. For index determination Use one of the following stains: A (grass on Cotton, EMPA 164), B (wholemeal / soot on cotton, 10N), C (blood / milk on cotton, C-5 (044)).

This Test material is preferred with the surfactant-containing solution a dissolved detergent, d. H. a wash liquor, washed for 60 minutes at a temperature of 40 ° C, in the presence and absence of the respective protease. For investigation The proteolysis index can in principle be different Detergent formulations are used, primarily important is the presence of a surfactant. If a detergent recipe is used, the dosage is 5.9 g of the detergent per Liter of wash liquor. The detergent or the surfactant-containing solution may, but does not necessarily have to contain bleach. The one to use Water is city water with a water hardness of about 16 ° German Hardness. A particularly suitable detergent formulation is compounded as follows (all figures in weight percent): 10% linear alkyl benzene sulfonate (sodium salt), 1.5% C12-C18 fatty alcohol sulfate (Sodium salt), 2.0% C12-C18 fatty alcohol with 7 EO, 20% sodium carbonate, 6.5% sodium bicarbonate, 4.0% amorphous sodium disilicate, 17 % Sodium Carbonate Peroxohydrate, 4.0% TAED, 3.0% Polyacrylate, 1.0% Carboxymethyl cellulose, 1.0% phosphonate, 25% sodium sulfate, remainder: Foam inhibitors, optical brightener, fragrances, possibly water.

Generally the surfactant is selected from those described above Surfactants. The surfactant is in proportions of 5 wt .-% to 50 wt .-%, in particular from 8% to 30% by weight, contained in the solution. The measurement of the degree of whiteness is done on a previously with a white standard calibrated spectrometer in a standard, Wei se known in the art. The Proteolyseindex is then defined as the quotient of the measured value of the degree of whiteness the surfactant-containing solution with a protease according to the invention from Xanthomonas and the measurement of the whiteness of the surfactant-containing solution without an inventive Protease from Xanthomonas over a pH range of at least 0.5, and more preferably from 0.75, from 1, from 1.25, of 1.5, 1.75 and 2 pH units. This pH range is preferably in a neutral or alkaline pH range, d. H. the smallest pH in this pH range is greater or equal to pH 7.

Of the Proteolysis index therefore expresses that an inventive Protease over a wider pH range a corresponding cleaning performance, d. H. Catalytic power, provides what they are for use in detergents and cleaners makes it particularly advantageous.

In a preferred embodiment of this subject invention is the protease naturally present in a bacterium, which is selected from the group consisting of Xanthomonas albilineans, Xanthomonas alfalfae, Xanthomonas alfalfae subsp. alfalfa, xanthomonas alfalfae subsp. citrumelonis, Xanthomonas ampelina (Xylophilus ampelinus), Xanthomonas arboricola, Xanthomonas axonopodis, Xanthomonas bromi, Xanthomonas campestris, Xanthomonas cassavae, Xanthomonas citri, Xanthomonas citri subsp. citri, Xanthomonas citri subsp. malvacearum, Xanthomonas codiaei, Xanthomonas cucurbitae, Xanthomonas cynarae, Xanthomonas euvesicatoria, Xanthomonas fragariae, Xanthomonas fuscans, Xanthomonas fuscans subsp. aurantifolii, Xanthomonas fuscans subsp. fuscans, Xanthomonas gardneri, Xanthomonas hortorum, Xanthomonas hyacinthi, Xanthomonas melonis, Xanthomonas oryzae, Xanthomonas perforans, Xanthomonas phaseoli, Xanthomonas pisi, Xanthomonas populi, Xanthomonas sacchari, Xanthomonas theicola, Xanthomonas translucens, Xanthomonas vasicola, Xanthomonas vesicatoria. Especially preferred is the protease naturally present in a bacterium, which is selected from the group consisting of Xanthomonas axonopodis DSM 3585, Xanthomonas axonopodis pvar. begoniae DSM 50850, Xanthomonas axonopodis pvar. malvacearum DSM 1220, Xanthomonas campestris DSM 1050 (NCPPB 1929), Xanthomonas campestris DSM 19000, Xanthomonas campestris pvar. campestris DSM 3586T, Xanthomonas campestris pvar. pelargonii DSM 50857.

Under For the purposes of the present application, an enzyme is a protein to understand that has a certain biocatalytic function. Protease in the sense of the present application is an enzyme which catalyzes the hydrolysis of peptide bonds and thereby be able to cleave peptides or proteins.

One Protein is one of the natural within the meaning of the present application Amino acids composed, largely linearly structured, to perform its function mostly three-dimensional structure accepting polymer, d. H. a polypeptide to understand. A peptide In the present application, the proteinogenic, naturally occurring L-amino acids with the internationally commonly used 1- and 3-letter codes. numerous Proteins are called preproteins, ie together formed with a signal peptide. Below that is the N-terminal Part of the protein whose function is mostly to the discharge of the formed protein from the producing cell into the periplasm or the surrounding medium and / or its correct one To ensure folding. Subsequently, the Signal peptide under natural conditions by a signal peptidase split off from the remaining protein, so this is its actual catalytic activity without the initial ones Exerts N-terminal amino acids. Proproteins are inactive precursors of proteins. Their forerunner with signal sequence are referred to as pre-pro-proteins. For technical applications are due to their enzymatic Activity the valves, d. H. mature peptides, d. H. the After their preparation, enzymes were processed in relation to the Preproteins preferred. The proteins can be from the cells producing them after the production of the polypeptide chain be modified, for example, by attaching Sugar molecules, formylations, aminations, etc. Such Modifications are referred to as post-translational modifications. These post translational modifications may need however, do not exert any influence on the function of the protein. A fragment is part of a protein. In this sense also a mature peptide is a fragment relative to the corresponding one Preprotein. A derivative is a protein that binds to at least an amino acid has a chemical modification. In this sense, posttranslational modified proteins Derivatives. For the purposes of the present invention, the term "protein" therefore stand for all enzymes, proteins, fragments and derivatives, unless explicitly addressed as such.

By Comparison with known enzymes, for example, in general accessible databases From the amino acid or nucleotide sequence the enzymatic Activity of a considered enzyme infer. This can through other areas of the protein that are not at the actual Reaction involved, qualitatively or quantitatively modified become. This could be, for example, the enzyme stability, the activity, the reaction conditions or the substrate specificity affect.

Such a comparison is accomplished by associating similar sequences in the nucleotide or amino acid sequences of the proteins of interest. This is called homologization. A tabular assignment of the respective positions is referred to as alignment. In the analysis of nucleotide sequences, in turn, both complementary strands and all three possible reading frames are to be taken into account; as well as the degeneracy of the genetic code and the organism-specific use of codons (codon usage). Meanwhile, alignments are created using computer programs, such as the algorithms FASTA or BLAST; This procedure is for example by DJ Lipman and WR Pearson (1985) in Science, Vol. 227, pp. 1435-1441 described. A summary of all matching positions in the compared sequences is called a consensus sequence.

Such a comparison also allows a statement about the similarity or homology of the compared sequences to each other. This is represented in percent identity, that is the proportion of identical nucleotides or amino acid residues at the same or in an alignment corresponding positions. A broader concept of homology includes the conserved amino acid substitutions in this value. It then speaks of percent similarity. Such statements can be made about whole proteins or genes or only over individual areas. In this way, a sequence space can be defined. Often this is related to one or more specific starting sequences and defined by a percentage of the identity or homology score. A sequence is then part of the sequence space if the comparison with an output sequence gives a higher identity or homology value in comparison with the identity or homology value defining the sequence space. Furthermore, however, it is also possible to define the sequence space by specifying those molecules which are to be contained in it. By specifying which molecules is to comprise the sequence space, it is possible to obtain or isolate the molecules, to determine the sequences (in proteins, preferably the amino acid sequence before the nucleic acid sequence) of these molecules, to assign them in a sequence comparison, and thus the To determine the sequence space defining identity or homology value. All of these procedures necessary for this are state of the art and the expert in the field of enzyme technology long known.

• a) Proteasen, die dadurch gekennzeichnet sind, dass sie in einem Bakterium der Gattung Xanthomonas natürlicherweise vorhanden sind und sie in einer wässrigen Lösung in Anwesenheit von mindestens einem Tensid einen Proteolyseindex von mindestens 1,1 aufweisen und/oder
• b) Proteasen, die natürlicherweise vorhanden sind in einem Bakterium, welches aus einer der vorstehend genannten Gruppen ausgewählt ist.

In the context of the present invention, the proteases that are naturally present in a bacterium of the genus Xanthomonas define such a sequence space. Further sequence spaces are defined by

• a) proteases characterized in that they are naturally present in a bacterium of the genus Xanthomonas and have a proteolysis index of at least 1.1 in an aqueous solution in the presence of at least one surfactant and / or
• b) proteases which are naturally present in a bacterium which is selected from one of the abovementioned groups.

All Proteases of said sequence spaces are within the scope of protection of the present invention included.

homologous Areas of different proteins are represented by correspondences, especially matches, in the amino acid sequence Are defined. These can also have an identical function to be marked. She goes to complete identities in the smallest areas, so-called boxes, which have only a few amino acids include and mostly for the overall activity one perform essential functions. Among the functions of homologous areas are the smallest sub-functions of the entire Protein function understood, such as the formation of individual hydrogen bonds to Complexation of a substrate or transition complex.

proteases or enzymes in general can be prepared by various methods, z. B. targeted genetic modification by mutagenesis, evolved and for specific purposes or in terms of special properties, for example with regard to their catalytic Activity, its stability under certain conditions, etc., to be optimized. Such enzyme variants are made from a Precursor enzyme, for example, a wild-type enzyme by Alteration of the amino acid sequence generated. The Change of the amino acid sequence is preferably carried out by mutations, with amino acid substitutions, deletions, Insertions or combinations thereof may be made. The introduction of such mutations into proteins is state of the art and those skilled in the art of enzyme technology known. It is furthermore generally known from the prior art that advantageous properties of individual mutations, for. B. single point mutations, can complement. A with respect to certain properties already optimized protease, for example, in terms of their stability Surfactants or other components, according to the invention additionally be further developed.

proteins can be about the reaction with an antiserum or a particular antibody to groups more immunologically related Proteins are summarized. The members A group is characterized by being the same, by one Antibodies recognized to have antigenic determinant. With included in the scope are therefore also proteases, which are characterized by having at least one and increasingly preferably two, three or four matching antigens Determinants with a protease according to the invention exhibit.

a Another object of the invention is nucleic acid molecules, that for a protease according to the invention encode as well as vectors containing such a nucleic acid.

Under Nucleic acids are within the meaning of the present application naturally constructed from nucleotides as information carriers serving for the linear molecules Code amino acid sequence in proteins or enzymes. she can be as a single strand, as one to this single strand complementary single strand or as a double strand. As the naturally more permanent information carrier the nucleic acid DNA for molecular biological Works preferred. In contrast, for the Realization of the invention in natural environment, such as for example, in an expressing cell, an RNA is formed, which is why essential RNA molecules also embodiments of the present invention.

For DNA, consider the sequences of both complementary strands in all three possible reading frames. Further, it should be appreciated that different codon triplets may encode the same amino acids so that a particular amino acid sequence may be derived from a plurality of different and possibly low identity nucleotide sequences, which is referred to as the degeneracy of the genetic code. In addition, different organisms submerge Divide in the use of these codons. For these reasons, both amino acid sequences and nucleotide sequences must be included in the consideration of the scope. Therefore, all nucleotide sequences are included in the invention which can encode an enzyme as described above. The person skilled in the art is able to determine these nucleotide sequences unequivocally since, despite the degeneracy of the genetic code, individual codons can be assigned to defined amino acids. The information unit corresponding to a protein is also referred to as gene within the meaning of the present application.

The The present invention also encompasses the production of recombinant Proteins. These include all according to the invention to understand genetic engineering or microbiological processes that based on the fact that the genes are of interest to the Proteins into a host cell suitable for production, which is a non-human cell, introduced and from this be transcribed and translated. Suitably, the Introduction of the relevant genes via vectors, in particular Expression vectors; but also about those that cause that the gene of interest in the host cell into an already existing inserted genetic element such as the chromosome or other vectors can be. The functional unit of gene and promoter and eventual other genetic elements is inventively as Expression cassette called. She has to for it but not necessarily as a physical entity.

A person skilled in the art is now able to use well-known methods such as, for example, chemical synthesis or polymerase chain reaction (PCR) in conjunction with standard molecular biological and / or proteinchemical methods, using known DNA and / or amino acid sequences, the appropriate nucleic acids up to to produce complete genes. Such methods are for example Sambrook, J., Fritsch, EF and Maniatis, T. 2001. Molecular cloning: a laboratory manual, 3rd Edition Cold Spring Laboratory Press. known.

Under For the purposes of the present invention, vectors are made from nucleic acids existing elements understood as the characteristic nucleic acid region contain a gene of interest. You can do this in a species, preferably a microorganism, or a cell line several generations or cell divisions away as a stable genetic Establish element. Vectors are especially in use in bacteria special plasmids, so circular genetic elements. On the one hand, one differentiates between such vectors, which the Storage and thus to a certain extent also the genetic engineering Work, the so-called cloning vectors, and on the other hand those who perform the function, the gene of interest to realize in the host cell, that is, the expression of the protein in question. These vectors are termed expression vectors.

in the The present invention relates to the nucleic acid suitably cloned into a vector. Vectors can of bacterial plasmids, viruses or bacteriophages derive or predominantly synthetic vectors or plasmids be with elements of different origin. With the others, respectively existing genetic elements can vectors themselves in the respective host cells over several generations away as stable units. It is in the sense of doing so irrelevant to the invention, whether they are extrachomosomal as their own Establish units or integrate into a chromosome. The selection a vector, for example, based on the achievable copy number, the available selection systems, including especially antibiotic resistance, or on the basis of cultivability the host cells into which the vector is to be introduced.

The Vectors form suitable starting points for molecular biology and biochemical studies of the gene or its associated Protein and for further developments and also for the amplification and production of the invention Proteins. They represent insofar embodiments of the present Invention, provided they nucleic acids according to the invention full. H. a nucleic acid is present in the vector is that for an inventive enzyme coded.

a Another object of the invention thus represent vectors that contain at least one nucleic acid molecule, which for a protease according to the invention coded. In a specific embodiment of the invention the vector is characterized in that the vector is a cloning vector is. These cloning vectors are useful in addition to storage, the biological amplification or selection of the subject of interest Gene for the characterization of the gene in question, about creating a restriction map or sequencing. Further provide a transportable and storable form of the claimed DNA. They are also preferred starting points for molecular biology techniques that are not attached to cells are bound, such as the polymerase chain reaction.

Prefers the vector is an expression vector. Expression vectors are chemical Similar to the cloning vectors, but differ in those subsequences that enable them to enter the for the production of proteins optimized host cells or host organisms to replicate and there the gene contained to bring to expression. Preferred embodiments are expression vectors that themselves are necessary for expression carry genetic elements. For example, the expression becomes of promoters affecting the transcription of the Regulate gene. So expression can be influenced by the natural, originally done in front of this gene localized promoter, but also after a genetic fusion both by a on the Expression vector provided promoter of the host cell as well by a modified or a completely different promoter another organism or host cell. Especially can expression vectors about changes the culture conditions or addition of certain compounds, such as for example cell density or special factors, regulatable be. Expression vectors allow the associated Protein heterologous, so in another cell or host cell as the one from which it is naturally derived can, is produced. The cells can be quite too belonging to different organisms or from different ones Come from organisms. Also a homologous protein recovery from a the gene naturally expressing host cell an appropriate vector is within the scope of the present invention Invention. This may have the advantage that natural, translation related transformations be performed on the resulting protein as well as they would naturally occur.

A further embodiment represent expression systems, in which additional genes, such as those that be provided on other vectors that Influence production of proteins according to the invention. These may be modifying gene products or to those with the protein of the invention be cleaned together, about its enzymatic function to influence. These may be, for example, other proteins or enzymes to inhibitors or to act on such elements influence the interaction with different substrates.

One Another object of the invention is a non-human host cell, which contains a protein of the invention or which can be stimulated to produce them, preferably below Use of an expression vector. The in vivo synthesis of a Enzyme, that is by living cells, requires the transfer of the associated gene in a host cell, their so-called transformation. As host cells are in principle all cells, that is prokaryotic or eukaryotic cells. Preferred are such Host cells that are genetically beneficial to handle, what for example, the transformation with the expression vector and its stable establishment, for example unicellular mushrooms or bacteria. In addition, preferred host cells are characterized a good microbiological and biotechnological handling out. This concerns, for example, easy cultivability, high Growth rates, low requirements for fermentation media and good production and secretion rates for foreign proteins. Often, from the wealth of different prior art systems the optimal expression systems for the individual case experimentally be determined. Each protein of the invention In principle, it can be made up of a large number of host cells be won. in particular, such host cells are preferred which are characterized in that they are after transformation with one of the vectors described above. there For example, they may be cloning vectors that for storage and / or modification, for example in a bacterial strain or another host cell according to the invention introduced have been. Such steps are in storage and further development genetic elements. Because of these Host cells transfer the relevant genetic elements into subsequent, immediately transferred to the expression of suitable host cells can be, are the same with the previous ones Transformation products to realizations of this subject invention.

preferred Embodiments illustrate such host cells that are due to genetic regulatory elements, for example, on the expression vector be made available, but also from the outset may be present in these cells, in their activity are adjustable. For example, by controlled addition of chemical compounds that serve as activators, by amendment the cultivation conditions or when a certain Cell density, these can be excited for expression. This allows an economical production of the invention Proteins.

Preferred host cells are prokaryotic or bacterial cells. As a rule, bacteria are distinguished from eukaryotes by shorter generation times and lower demands on culturing conditions. As a result, cost-effective methods for obtaining proteins according to the invention can be established. Gram-negative bacteria such as Escherichia coli (E. coli) secrete a variety of proteins into the periplasmic space, the compartment between the two the cells enclosing membranes. This can be advantageous for special applications. In contrast, Gram-positive bacteria, such as, for example, Bacilli or Actinomycetes or other representatives of the Actinomycetales, have no outer membrane, so that secreted proteins are released into the nutrient medium surrounding the cells, from which the expressed proteins according to the invention can be directly purified. Preferably, the host cell secretes the protein of the invention into the surrounding medium.

• a) der Gruppe der Gattungen von Escherichia, Bacillus und Arthrobacter, Streptomyces, Stenotrophomonas, Xanthomonas und Pseudomonas oder
• b) der Gruppe von Escherichia coli, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus alcalophilus und Arthrobacter oxidans, Streptomyces lividans, Streptomyces coelicolor und Stenotrophomonas maltophilia.

In a preferred embodiment, the host cell according to the invention is characterized in that it is a bacterium, in particular one which is selected from

• a) of the genera Escherichia, Bacillus and Arthrobacter, Streptomyces, Stenotrophomonas, Xanthomonas and Pseudomonas or
• b) the group of Escherichia coli, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus alcalophilus and Arthrobacter oxidans, Streptomyces lividans, Streptomyces coelicolor and Stenotrophomonas maltophilia.

Especially suitable host cells are bacteria of the genus Xanthomonas and most preferably one mentioned in Table 1 Xanthomonas species or a named in Table 1 Xanthomonas strain.

The Host cells can meet their requirements the cultural conditions have changed, others or additional Have selection markers or other or additional Express proteins. It may in particular be such host cells acting in addition to the protein produced according to the invention even more, especially economically interesting proteins express.

The However, the host cell can also be a eukaryotic cell that is characterized characterized in that it has a nucleus. Another one The invention therefore provides a host cell which is characterized characterized in that it has a nucleus. In contrast to prokaryotic cells, eukaryotic cells are able to modify the protein formed posttranslationally. Examples for it are mushrooms like Actinomyceten or yeasts like Saccharomyces or Kluyveromyces. This can be particularly advantageous, for example when the proteins are specific in the context of their synthesis To undergo modifications that allow such systems. These include, for example, the binding of low molecular weight Compounds such as membrane anchors or oligosaccharides.

The Host cells according to the invention are in themselves cultivated and fermented in a known manner, for example in discontinuous or continuous systems. In the first case will be a suitable one Nutrient medium inoculated with the host cells and the product after an experimentally determined period of time from the medium harvested. Continuous fermentations are characterized by reaching a steady state in which over a relatively long-term cells partly die off but regrow and simultaneously remove product from the medium can be.

fermentation process are known per se from the prior art and represent the actual large-scale technical Production step, usually followed by a suitable one Purification method of the product produced, for example of the recombinant protein. Here are the for the manufacturing methods used, for the host cells and / or the proteins to be produced each have optimal conditions based on the previously optimized culture conditions of the relevant Strains according to the knowledge of the skilled person, for example Fermentation volume, media composition, oxygenation or Stirrer speed to be determined experimentally.

Fermentation process, which are characterized in that the fermentation over An inflow strategy is also implemented into consideration. Here are the media components, by the continuous cultivation are consumed, fed; One also speaks of a feeding strategy. hereby can bring significant increases in both the Cell density, as well as in the dry biomass and / or especially the Activity of the protein of interest can be achieved.

Analogous In addition, the fermentation can also be designed so that unwanted Metabolites filtered out or by adding buffer or each appropriate counterions are neutralized.

The produced protein can be harvested subsequently from the fermentation medium. This fermentation process is preferred over dry matter product processing, but requires the provision of suitable secretion markers and transport systems. Without secretion u. U. the purification of the protein from the cell mass required, and various methods are known, such as precipitation z. B by ammonium sulfate or ethanol, or the chromatographic purification, if necessary to homogeneity. However, the majority of the described technical methods should manage with an enriched, stabilized preparation.

All Already executed above elements can be used to process be combined to proteins of the invention manufacture. The optimal procedure is for each concrete To determine and optimize an individual case experimentally. This Procedure is in the routine area of one with enzyme production trusted professional.

a independent subject of the invention thus also provide Process for the preparation of an inventive Protease

To Every method that comes to making a top belongs described protease invention suitable or that is the receipt of an inventive Protease allows. These include, for example also chemical synthesis methods. In contrast, preferred However, all are well established in the art, above in individual Aspects already discussed molecular biological, microbiological, or biotechnological production process.

• a) Kultivieren einer entsprechenden Wirtszelle
• b) Isolieren der Protease aus den kultivierten Wirtszellen und/oder aus dem die Wirtszellen umge benden Medium
• c) optional Reinigen der isolierten Protease

An inventively preferred method for producing a protease comprises the method steps

• a) culturing a corresponding host cell
• b) isolating the protease from the cultured host cells and / or from the medium surrounding the host cells
• c) optionally purifying the isolated protease

Preferably these are procedures that are made using a previously designated nucleic acid, preferably using a as described above, and more preferably using a previously designated, advantageously genetically modified host cell done. Because this is the corresponding preferred genetic information in a microbiologically usable form made available. The central aspect here is that the host cell is capable of carrying out a method according to the invention Expressing protease. This can be achieved by using a for nucleic acid encoding such a protease into the host cell was introduced. Preferably, this nucleic acid became introduced into the host cell as part of an expression vector, so that the host cell is able to express the protease is. Furthermore, it is possible that the nucleotide sequence of the Vector in one or more codons to the codon usage of the host strain has been adjusted.

A Embodiment of an inventive Manufacturing process can be based on the associated Nucleic acid sequence also a cell-free expression system be in which the protein biosynthesis is understood in vitro.

a Another subject of the invention provide methods for the purification of Textiles or hard surfaces where at least in one of the method steps an inventive Protease is used. The procedure for cleaning textiles or hard surfaces is thus characterized that in at least one method step, an inventive Protease is proteolytically active.

this includes fall both manual and mechanical procedures, whereby machine Method because of their more precise controllability, what For example, the amounts used and exposure times, preferably are.

method for the cleaning of textiles are generally characterized from that in several process steps different cleaning active Substances applied to the items to be cleaned and after the exposure time be washed off, or that the items to be cleaned in any other way with a detergent or a solution of this agent is treated. The same applies to procedures for cleaning of all materials other than textiles, which are termed "hard Surfaces "are summarized. All wash or Cleaning methods may be used in at least one of the method steps enriched by a protease according to the invention and then provide embodiments of the present invention represents.

In the methods of the invention, the protease is added in an amount of from 40 μg to 4 g, preferably from 50 μg to 3 g, more preferably from 100 μg to 2 g and most preferably from 200 μg to 1 g used per application.

There preferred enzymes of the invention naturally already have a protein-dissolving activity and they also unfold in media, which otherwise no cleaning power own, such as in bare buffer, can single sub-step of such a method for machine Cleansing of textiles consist of that in addition to stabilizing Compounds, salts or buffer substances as sole cleaning active Component applied an enzyme of the invention becomes. This constitutes a further embodiment of this subject of the invention represents.

a Another own subject of the invention is the use of a Protease of the invention described above for the removal of protease-sensitive stains on textiles or hard surfaces, d. H. for cleaning textiles or hard surfaces.

Because Proteases of the invention can, in particular because of their properties described above, used be proteinaceous of textiles or hard surfaces Eliminate impurities. Embodiments of this use For example, hand wash, manual removal of patches of textiles or of hard surfaces or the machine cleaning of textiles or hard surfaces In a preferred embodiment of this use are the respective proteases of the invention provided according to one of the above recipes.

In a use according to the invention becomes the protease in an amount of 40 μg to 4 g, preferably 50 μg to 3 g, more preferably from 100 μg to 2 g and whole more preferably used from 200 μg to 1 g per application.

The The following examples further illustrate the invention, without but to limit it to:

Example 1: Cultivation of the strains and screening for proteases

The Cultivation of the bacteria of the genus Xanthomonas took place in nutrient Broth medium (peptone 5.0 g / l and meat extract 3.0 g / l) Shake overnight at 30 ° C with 180 revolutions per minute. The bacteria thus obtained were based on milk powder Agar plates (peptone 5.0 g / l and meat extract 3.0 g / l and agar 15.0 g / l and 2% skimmed milk powder) and plated for 48 hours Incubated at 30 ° C. On the basis of a Klärungshofes (Lysehof) around the respective bacterial colony showed a proteolytic Activity of the bacteria. They were cultivated again, such that the culture medium is the protease formed by the bacteria and thus containing proteolytic activity.

On this way became the proteolytic activity of bacteria of the Xanthomonas strains Xanthomonas axonopodis DSM 3585, Xanthomonas axonopodis pvar. Begoniae DSM 50850, Xanthomonas axonopodis pvar. malvacearum DSM 1220, Xanthomonas campestris DSM 1050 (NCPPB 1929), Xanthomonas campestris DSM 19000, Xanthomonas campestris pvar. campestris DSM 3586T and Xanthomonas campestris pvar. pelargonii DSM 50857 received.

Example 2: Determination of the Washing Performance when used in commercial detergent formulations

It Standardized soiled textiles were used by the Federal Materials Testing and Testing Institute, St. Gallen, Switzerland (EMPA), or the laundry research institute, Krefeld, had been obtained. The following stains were and textiles used: A (grass on cotton, EMPA 164), B (whole / soot on Cotton, 10N), C (blood / milk on cotton, C-5 (044)).

With This test material has various detergent formulations, the differed by the particular protease contained on their Washing performance examined. That's what the approaches became washed for 60 minutes at a temperature of 40 ° C. The dosage was 5.9 g of the detergent per liter of wash liquor. It was made with city water with a water hardness of about 16 ° German hardness washed in a pH range between pH 8 and pH 9. The detergent formulation was composed as follows (all figures in weight percent): 10% linear alkyl benzene sulfonate (Sodium salt), 1.5% C12-C18 fatty alcohol sulfate (sodium salt), 2.0% C12-C18 fatty alcohol with 7 EO, 20% sodium carbonate, 6.5% sodium bicarbonate, 4.0% amorphous sodium disilicate, 17% sodium carbonate peroxohydrate, 4.0% TAED, 3.0% polyacrylate, 1.0% carboxymethylcellulose, 1.0% phosphonate, 25% sodium sulfate, balance: foam inhibitors, optical brightener, Fragrances, if necessary. Water.

The Detergent formulation was used for the different test series identical to the various proteases from the Xanthomonas strains. The used protease activity was either 5 or 10 PE (protease units) per ml wash liquor. The same procedure was followed with reference formulas, provided they were Contained proteases.

To washing was the whiteness of the washed textiles, d. H. the brightening of soiling, measured. The measurement was made on a spectrometer Minolta CM508d (illuminant D65, 10 °). The Device was previously with a supplied white standard calibrated.

The Detergents with Xanthomonas proteases had a high washing performance on. In particular, these formulations had a better washing performance on the soiled soils as the comparative formulations used. The proteases from Xanthomonas therefore have a proteolysis index of at least 1.1. This was determined as the quotient of obtained results in a washing process with a detergent containing a protease and a parallel performed Control wash with the detergent without protease.

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

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Claims (18)

Washing or cleaning agent comprising a protease, in a bacterium of the genus Xanthomonas naturally is present and at least one other detergent ingredient. Washing or cleaning agent according to one of the claims 1, characterized in that the further detergent ingredient is selected from the group consisting of builder substances, Surface-active surfactants, bleaching agents based on organic and / or inorganic peroxygen compounds, bleach activators, water-miscible organic solvents, enzymes, sequestering agents, Electrolytes, pH regulators and other excipients such as optical Brightener, grayness inhibitors, foam regulators and color and Perfumes and combinations thereof. Washing or cleaning agent according to one of the claims 1 or 2, characterized in that it is in solid form, in particular as free-flowing powder with a bulk density from 300 g / l to 1200 g / l, in particular 500 g / l to 900 g / l. Washing or cleaning agent according to one of the claims 1 to 2, characterized in that it is in liquid, gel or pasty form is present. Washing or cleaning agent according to one of the claims 1 to 4, characterized in that the protease with a at Room temperature and / or in the absence of water for the protease impermeable substance envelops is. Washing or cleaning agent according to one of the claims 1 to 5, characterized in that there is at least one other Enzyme, in particular a protease, amylase, cellulase, hemicellulase, mannanase, Tannase, xylanase, xanthanase, β-glucosidase, carrageenase, Oxidase, oxidoreductase, lipase or combinations thereof. Use of a washing or cleaning agent after any one of claims 1 to 6 for the removal of protease-sensitive Stains on textiles or hard surfaces. Process for cleaning textiles or hard Surfaces, characterized in that in at least a process step, a washing or cleaning agent after a of claims 1 to 6 is applied. Protease, characterized in that it is in a Bacterium of the genus Xanthomonas naturally present is and they are in an aqueous solution in the presence of at least one surfactant has a proteolysis index of at least 1.1 has. Nucleic acid molecule, encoding for a protease according to claim 9. Vector containing a nucleic acid Claim 10. Non-human host cell that is a protease according to claim 9 or a fragment thereof or the can be excited to their preparation, preferably using a vector according to claim 11. Host cell according to claim 12, characterized in that that it has a nucleus. Host cell according to claim 12, characterized in that that she is a bacterium. Host cell according to claim 14, characterized in that that the bacterium is selected from a) the group the genera of Escherichia, Bacillus and Arthrobacter, Streptomyces, Stenotrophomonas, Xanthomonas and Pseudomonas or b) the group of Escherichia coli, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus alcalophilus and Arthrobacter oxidans, Streptomyces lividans, Streptomyces coelicolor and Stenotrophomonas maltophilia. A method of producing a protease according to claim 9 comprising the steps of a) culturing a host cell according to any one of claims 12 to 15 b) isolating the protease from the cultured host cells and / or from the medium surrounding the host cells c) optionally purifying the isolated protease Process for cleaning textiles or hard Surfaces, characterized in that in at least a process step a protease according to claim 9 proteolytically is active. Use of a protease according to claim 9 for purification of textiles or hard surfaces.